Diva-Fit: A step-by-step manual for generating high-resolution graphs and histogram overlays of flow cytometry data obtained with FACSDiva software

In recent years, flow cytometry has been revolutionized via the introduction of digital data acquisition and analysis tools that facilitate simultaneous investigation of ten or more parameters. At the same time, some data presentation tools offered by commercial suppliers have remained surprisingly “antiquated”. This leads to the ironic fact that high-quality data is often represented by poor-quality illustrations: namely pixelated plots. In particular, data obtained using FACSDiva software is frequently exported into figures as low-resolution pixel graphics resulting in low-quality images. Additionally, even the newest version, Diva_6.1.2, is still unable to generate histogram overlays, a popular and convincing tool for direct data comparison. We hereby present an easy and down-to-earth Diva-Figure-improvement toolbox (Diva-Fit), which facilitates the generation of high-resolution graphs based on data acquired with FACSDiva software. Moreover, Diva-Fit allows the easy removal of unwanted quadrant labels without impairing the quality of FACS plots. Finally, we show that Diva-Fit supports straightforward composition of histogram overlays. All software tools necessary are freely available. We believe that the proposed toolbox may be very useful for many researchers working with flow cytometry.